Layered structures of fiber cell wall of Korean red pine (Pinus densiflora) were investigated by confocal reflection microscopy (CRM). CRM micrographs revealed detailed structures of the fiber cell wall such as S1, S2, and S3 layers as well as transition layers (S12 and S23 layers), which are present between the S1, S2, and S3 layers. Microfibril angle (MFA) measurement was possible for the S2 and S3 layer in the cell wall. The experimental results suggest that CRM is a versatile microscopic method for investigation of layered structures and MFA measurement in individual sub layer of the tracheid cell wall.